The HIV 5' Gag Region Displays a Specific Nucleotide Bias Regulating Viral Splicing and Infectivity

Viruses. 2021 May 27;13(6):997. doi: 10.3390/v13060997.


Alternative splicing and the expression of intron-containing mRNAs is one hallmark of HIV gene expression. To facilitate the otherwise hampered nuclear export of non-fully processed mRNAs, HIV encodes the Rev protein, which recognizes its intronic response element and fuels the HIV RNAs into the CRM-1-dependent nuclear protein export pathway. Both alternative splicing and Rev-dependency are regulated by the primary HIV RNA sequence. Here, we show that these processes are extremely sensitive to sequence alterations in the 5'coding region of the HIV genomic RNA. Increasing the GC content by insertion of either GFP or silent mutations activates a cryptic splice donor site in gag, entirely deregulates the viral splicing pattern, and lowers infectivity. Interestingly, an adaptation of the inserted GFP sequence toward an HIV-like nucleotide bias reversed these phenotypes completely. Of note, the adaptation yielded completely different primary sequences although encoding the same amino acids. Thus, the phenotypes solely depend on the nucleotide composition of the two GFP versions. This is a strong indication of an HIV-specific mRNP code in the 5' gag region wherein the primary RNA sequence bias creates motifs for RNA-binding proteins and controls the fate of the HIV-RNA in terms of viral gene expression and infectivity.

Keywords: HIV gene expression; U1 snRNP; alternative splicing; mRNP code.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing
  • HEK293 Cells
  • HIV Infections / virology
  • HIV-1 / genetics*
  • HIV-1 / pathogenicity
  • Humans
  • RNA Splice Sites
  • RNA Splicing*
  • RNA, Messenger
  • RNA, Viral / genetics
  • Virus Replication / genetics*
  • gag Gene Products, Human Immunodeficiency Virus / genetics*


  • RNA Splice Sites
  • RNA, Messenger
  • RNA, Viral
  • gag Gene Products, Human Immunodeficiency Virus